A dual-color luciferase assay system reveals circadian resetting of cultured fibroblasts by co-cultured adrenal glands
In mammals, circadian rhythms of various organs and tissues are synchronized by pacemaker neurons in the suprachiasmatic nucleus (SCN) of the hypothalamus. Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in the...
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| description | In mammals, circadian rhythms of various organs and tissues are synchronized by pacemaker neurons in the suprachiasmatic nucleus (SCN) of the hypothalamus. Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in their plasma concentrations; however, whether organs outside the SCN can serve as master synchronizers to entrain circadian rhythms in target tissues is not well understood. To further delineate the function of the adrenal glands and the interactions of circadian rhythms in putative master synchronizing organs and their target tissues, here we report a simple co-culture system using a dual-color luciferase assay to monitor circadian rhythms separately in various explanted tissues and fibroblasts. In this system, circadian rhythms of organs and target cells were simultaneously tracked by the green-emitting beetle luciferase from Pyrearinus termitilluminans (ELuc) and the red-emitting beetle luciferase from Phrixothrix hirtus (SLR), respectively. We obtained tissues from the adrenal glands, thyroid glands, and lungs of transgenic mice that expressed ELuc under control of the promoter from a canonical clock gene, mBmal1. The tissues were co-cultured with Rat-1 fibroblasts as representative target cells expressing SLR under control of the mBmal1 promoter. Amplitudes of the circadian rhythms of Rat-1 fibroblasts were potentiated when the fibroblasts were co-cultured with adrenal gland tissue, but not when co-cultured with thyroid gland or lung tissue. The phases of Rat-1 fibroblasts were reset by application of adrenal gland tissue, whereas the phases of adrenal gland tissue were not influenced by Rat-1 fibroblasts. Furthermore, the effect of the adrenal gland tissue on the fibroblasts was blocked by application of a glucocorticoid receptor (GR) antagonist. These results demonstrate that glucocorticoids are strong circadian synchronizers for fibroblasts and that this co-culture system is a useful tool to analyze humoral communication between different tissues or cell populations. |
| doi_str_mv | 10.1371/journal.pone.0037093 |
| format | Article |
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Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in their plasma concentrations; however, whether organs outside the SCN can serve as master synchronizers to entrain circadian rhythms in target tissues is not well understood. To further delineate the function of the adrenal glands and the interactions of circadian rhythms in putative master synchronizing organs and their target tissues, here we report a simple co-culture system using a dual-color luciferase assay to monitor circadian rhythms separately in various explanted tissues and fibroblasts. In this system, circadian rhythms of organs and target cells were simultaneously tracked by the green-emitting beetle luciferase from Pyrearinus termitilluminans (ELuc) and the red-emitting beetle luciferase from Phrixothrix hirtus (SLR), respectively. We obtained tissues from the adrenal glands, thyroid glands, and lungs of transgenic mice that expressed ELuc under control of the promoter from a canonical clock gene, mBmal1. The tissues were co-cultured with Rat-1 fibroblasts as representative target cells expressing SLR under control of the mBmal1 promoter. Amplitudes of the circadian rhythms of Rat-1 fibroblasts were potentiated when the fibroblasts were co-cultured with adrenal gland tissue, but not when co-cultured with thyroid gland or lung tissue. The phases of Rat-1 fibroblasts were reset by application of adrenal gland tissue, whereas the phases of adrenal gland tissue were not influenced by Rat-1 fibroblasts. Furthermore, the effect of the adrenal gland tissue on the fibroblasts was blocked by application of a glucocorticoid receptor (GR) antagonist. These results demonstrate that glucocorticoids are strong circadian synchronizers for fibroblasts and that this co-culture system is a useful tool to analyze humoral communication between different tissues or cell populations.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0037093</identifier><identifier>PMID: 22615906</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adrenal glands ; Adrenal Glands - cytology ; Adrenal Glands - metabolism ; Adrenal Glands - physiology ; Animal tissues ; Animals ; Biology ; Brain research ; Cell Communication - physiology ; Cell culture ; Cell interactions ; Circadian rhythm ; Circadian Rhythm - genetics ; Circadian Rhythm - physiology ; Circadian rhythms ; Clock gene ; Coculture Techniques - methods ; Color ; Communications systems ; Cortical Synchronization - genetics ; Cortical Synchronization - physiology ; Fibroblasts ; Fibroblasts - cytology ; Fibroblasts - metabolism ; Fibroblasts - physiology ; Gene expression ; Genes ; Genetic engineering ; Glucocorticoids ; Homeostasis ; Hormones ; Hostages ; Hypothalamus ; Luciferase ; Luciferases - analysis ; Luciferases - metabolism ; Lungs ; Medical research ; Medicine ; Melatonin ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Organs ; Period Circadian Proteins - genetics ; Period Circadian Proteins - metabolism ; Promoter Regions, Genetic ; Rodents ; Signal Transduction ; Suprachiasmatic nucleus ; Suprachiasmatic Nucleus - metabolism ; Suprachiasmatic Nucleus - physiology ; Synchronism ; Synchronizers ; Thyroid ; Thyroid gland ; Tissues ; Tracking ; Transgenic mice</subject><ispartof>PloS one, 2012-05, Vol.7 (5), p.e37093-e37093</ispartof><rights>COPYRIGHT 2012 Public Library of Science</rights><rights>2012 Noguchi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Noguchi, Takako</au><au>Ikeda, Masaaki</au><au>Ohmiya, Yoshihiro</au><au>Nakajima, Yoshihiro</au><au>Foulkes, Nicholas S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A dual-color luciferase assay system reveals circadian resetting of cultured fibroblasts by co-cultured adrenal glands</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2012-05-15</date><risdate>2012</risdate><volume>7</volume><issue>5</issue><spage>e37093</spage><epage>e37093</epage><pages>e37093-e37093</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>In mammals, circadian rhythms of various organs and tissues are synchronized by pacemaker neurons in the suprachiasmatic nucleus (SCN) of the hypothalamus. Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in their plasma concentrations; however, whether organs outside the SCN can serve as master synchronizers to entrain circadian rhythms in target tissues is not well understood. To further delineate the function of the adrenal glands and the interactions of circadian rhythms in putative master synchronizing organs and their target tissues, here we report a simple co-culture system using a dual-color luciferase assay to monitor circadian rhythms separately in various explanted tissues and fibroblasts. In this system, circadian rhythms of organs and target cells were simultaneously tracked by the green-emitting beetle luciferase from Pyrearinus termitilluminans (ELuc) and the red-emitting beetle luciferase from Phrixothrix hirtus (SLR), respectively. We obtained tissues from the adrenal glands, thyroid glands, and lungs of transgenic mice that expressed ELuc under control of the promoter from a canonical clock gene, mBmal1. The tissues were co-cultured with Rat-1 fibroblasts as representative target cells expressing SLR under control of the mBmal1 promoter. Amplitudes of the circadian rhythms of Rat-1 fibroblasts were potentiated when the fibroblasts were co-cultured with adrenal gland tissue, but not when co-cultured with thyroid gland or lung tissue. The phases of Rat-1 fibroblasts were reset by application of adrenal gland tissue, whereas the phases of adrenal gland tissue were not influenced by Rat-1 fibroblasts. Furthermore, the effect of the adrenal gland tissue on the fibroblasts was blocked by application of a glucocorticoid receptor (GR) antagonist. These results demonstrate that glucocorticoids are strong circadian synchronizers for fibroblasts and that this co-culture system is a useful tool to analyze humoral communication between different tissues or cell populations.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>22615906</pmid><doi>10.1371/journal.pone.0037093</doi><tpages>e37093</tpages><oa>free_for_read</oa></addata></record> |
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| recordid | cdi_plos_journals_1344321677 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Public Library of Science (PLoS); PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Adrenal glands Adrenal Glands - cytology Adrenal Glands - metabolism Adrenal Glands - physiology Animal tissues Animals Biology Brain research Cell Communication - physiology Cell culture Cell interactions Circadian rhythm Circadian Rhythm - genetics Circadian Rhythm - physiology Circadian rhythms Clock gene Coculture Techniques - methods Color Communications systems Cortical Synchronization - genetics Cortical Synchronization - physiology Fibroblasts Fibroblasts - cytology Fibroblasts - metabolism Fibroblasts - physiology Gene expression Genes Genetic engineering Glucocorticoids Homeostasis Hormones Hostages Hypothalamus Luciferase Luciferases - analysis Luciferases - metabolism Lungs Medical research Medicine Melatonin Mice Mice, Inbred C57BL Mice, Transgenic Organs Period Circadian Proteins - genetics Period Circadian Proteins - metabolism Promoter Regions, Genetic Rodents Signal Transduction Suprachiasmatic nucleus Suprachiasmatic Nucleus - metabolism Suprachiasmatic Nucleus - physiology Synchronism Synchronizers Thyroid Thyroid gland Tissues Tracking Transgenic mice |
| title | A dual-color luciferase assay system reveals circadian resetting of cultured fibroblasts by co-cultured adrenal glands |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-23T22%3A48%3A31IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20dual-color%20luciferase%20assay%20system%20reveals%20circadian%20resetting%20of%20cultured%20fibroblasts%20by%20co-cultured%20adrenal%20glands&rft.jtitle=PloS%20one&rft.au=Noguchi,%20Takako&rft.date=2012-05-15&rft.volume=7&rft.issue=5&rft.spage=e37093&rft.epage=e37093&rft.pages=e37093-e37093&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0037093&rft_dat=%3Cgale_plos_%3EA477083229%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1344321677&rft_id=info:pmid/22615906&rft_galeid=A477083229&rft_doaj_id=oai_doaj_org_article_5fb6bf6c46a54b94a226848641db9d65&rfr_iscdi=true |